培养和诱导策略对重组大肠杆菌中BmR1抗原产生的影响。

Effects of feeding and induction strategy on the production of BmR1 antigen in recombinant E. coli.

作者信息

Norsyahida A, Rahmah N, Ahmad R M Y

机构信息

Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, 11800 Penang, Malaysia.

出版信息

Lett Appl Microbiol. 2009 Nov;49(5):544-50. doi: 10.1111/j.1472-765X.2009.02694.x.

DOI:10.1111/j.1472-765X.2009.02694.x

PMID:19832937

Abstract

AIM

To investigate the effects of feeding and induction strategies on the production of BmR1 recombinant antigen.

METHODS AND RESULTS

Fed-batch fermentation was studied with respect to the specific growth rate and mode of induction to assess the growth potential of the bacteria in a bioreactor and to produce high yield of BmR1 recombinant antigen. Cells were grown at a controlled specific growth rate (mu(set)) during pre-induction, followed by constant feeding postinduction. The highest biomass (24.3 g l(-1)) was obtained during fed-batch process operated at mu(set) of 0.15 h(-1), whereby lower mu(set) (0.075 h(-1)) gave the highest protein production (9.82 mg l(-1)). The yield of BmR1 was increased by 1.2-fold upon induction with 1 mmol l(-1) IPTG (isopropyl-beta-d-thiogalactoside) compared to using 5 mmol l(-1) and showed a further 3.5-fold increase when the culture was induced twice at the late log phase.

CONCLUSIONS

Combination of feeding at a lower mu(set) and twice induction with 1 mmol l(-1) IPTG yielded the best result of all variables tested, promising an improved method for BmR1 production.

SIGNIFICANCE AND IMPACT OF THE STUDY

This method can be used to increase the production scale of the BmR1 recombinant antigen to meet the increasing demand for Brugia Rapid(), a commercial diagnostic test for detection of brugian filariasis.

摘要

目的

研究补料和诱导策略对BmR1重组抗原生产的影响。

方法与结果

研究了分批补料发酵中特定生长速率和诱导方式，以评估生物反应器中细菌的生长潜力并高产BmR1重组抗原。细胞在诱导前以可控的特定生长速率（μ（设定值））生长，诱导后进行连续补料。在以0.15 h⁻¹的μ（设定值）运行的分批补料过程中获得了最高生物量（24.3 g l⁻¹），而较低的μ（设定值）（0.075 h⁻¹）则产生了最高的蛋白质产量（9.82 mg l⁻¹）。与使用5 mmol l⁻¹相比，用1 mmol l⁻¹异丙基-β-D-硫代半乳糖苷（IPTG）诱导时，BmR1的产量提高了1.2倍，当培养物在对数后期诱导两次时，产量进一步提高了3.5倍。